论文部分内容阅读
以雷林一号桉及尾叶桉为对象进行了原生质体分离体系建立研究，结果表明：雷林一号桉及尾叶桉胚性愈伤诱导的最适应培养基分别为：Ｂ５＋２，４－Ｄ２．０ｍｇ／Ｌ＋ＫＴ０．８ｍｇ／Ｌ及Ｂ５＋２，４－Ｄ２．０ｍｇ／Ｌ。胚性愈伤继代培养基为：Ｂ５＋２，４－Ｄ０．５～１．０ｍｇ／Ｌ＋ＫＴ０．５ｍｇ／Ｌ。黑暗下３～５天的预培养对愈伤组织上的褐化有明显的抑制作用。雷林一号桉及尾叶桉原生质体分离的最适酶液组合分别为：纤维素酶２％＋离析酶１％＋崩溃酶０．１％；纤维素酶２％＋离析酶１％＋崩溃酶０．０５％。愈伤组织在０．９ｍｏｌ／Ｌ蔗糖溶液中浸泡１ｈ，并于酶解后４ｈ更换一次酶液有利于原生质体产量与活力的提高。淡黄色松散愈伤组织原生质分离效果较其它质地的愈伤组织好。继代后８～１０天愈伤组织作原生质体的分离材料较其它年龄的愈伤组织合适。
The establishment of protoplast isolation system was studied on Eucalyptus urophylla and Eucalyptus urophylla. The results showed that the most suitable medium for inducing embryogenic callus of Eucalyptus urophylla and Eucalyptus urophylla were: B5 + 2,4- D2.0 mg / L + KT 0.8 mg / L and B5 + 2,4-D2.0 mg / L. Embryogenic callus subculture medium: B5 +2,4-D0.5 ~ 1.0mg / L + KT0.5mg / L. Pre-incubation of 3 to 5 days in the dark had a significant inhibitory effect on the callus browning. The optimal enzyme combinations for protoplast isolation of Eucalyptus urophylla and Eucalyptus urophylla were as follows: cellulase 2% + isozyme 1% + collage enzyme 0.1%; cellulase 2% + isozyme 1% Collapse enzyme 0.05%. Soaking callus in 0.9mol / L sucrose solution for 1h, and changing enzyme solution 4h after enzymolysis was beneficial to the increase of protoplast yield and vigor. Pale yellow loose callus protoplast isolation better than other callus texture. After subculture 8 to 10 days callus protoplast dissociation material more suitable than other age callus.